In this proposal we will build upon our 3D breast microtumor by 1) increasing the complexity of the breast microtumor, 2) developing non-small cell lung cancer and glioblastoma 3D microtumors, 3) validating the microtumor?s ability to predict drug response in vitro against the NCI60 and in vivo against patient derived xenografts (PDX), and 4) increasing our throughput with modifications to our 3DKUBETM. These microtumors will be used in the screening of anticancer agents, reducing clinical trial failure rates due to more relevant results, and to test patient derived tissues for real-time clinical decision making. Tissues will be developed with established cell lines. Following validation for morphology and function through lytic and non-lytic methods, patient derived samples will be incorporated and tested for their response to anticancer agents. As part of the validation, genetic, epigenetic, and miRNA based ?signatures? will be developed and compared across cell lines, 3D tissues, and in vivo samples to generate a biomarker panel and validate the relevancy of the microtumors. Validation against both the NCI60 and PDX models with approved and failed anticancer agents will further prove the predictive power of these models for drug development and real-time clinical decision making.